Surface plasmon resonance screening of synthetic peptides mimicking the immunodominant region of C-S8c1 foot-and-mouth disease virus

Abstract

The main antigenic site (site A) of foot-and-mouth disease virus (FMDV, strain C-S8c1) may be adequately reproduced by a 15-peptide with the amino acid sequence H-YTASARGDLAHLTTT-NH 2 (A15), corresponding to the residues 136±150 of the viral protein VP1. The eect of amino acid substitutions within A15 on its antigenicity towards monoclonal antibodies (MAb) raised against antigenic site A, has been studied by means of BIAcore technology, based on surface plasmon resonance (SPR). Although these antigenicities have previously been determined from enzyme-linked immunosorbent assays (ELISA), the SPRbased technique is superior in that it allows a fast and straightforward screening of antigens while simultaneously providing kinetic data of the antigen±antibody interaction. With a view to screening fairly large libraries of individual peptides, we have inverted the typical SPR experiment by immobilizing the MAb on the sensor surface and using peptides as soluble analytes. We report the validation of this approach through the screening of 44 site A peptides, with results generally in good agreement with the relative antigenicities previously determined by competition ELISA. #

Antigenicity modulation upon peptide cyclization: application to the GH loop of foot-and-mouth disease virus strain C1-Barcelona

“…As we have shown earlier, despite the fairly small size (ca. 1.6 kDa) of the peptides, reliable data can be obtained with this analysis configuration [10,11]. All the measurable interactions fitted to the pseudo-first-order Table 3 Affinity data for the interactions between mAbs SD6, 4C4 and 3E5, and peptides from the G-H loops of FMDV C-S8c1, C 1 -Brescia and C-S30 mAb Peptide a K A(kin1) , affinity from SPR kinetic analysis with immobilized mAb; K A(kin2) , affinity from SPR kinetic analysis with immobilized peptide; K i(sol.…”

Direct single-step surface plasmon resonance analysis of interactions between small peptides and immobilized monoclonal antibodies

“…ously described Gomes et al, 2000 . This preliminary study identified several key features for a suc-Ž .…”

“…In a previous report, we demonstrated that the SPR experimental set-up could be optimized to obtain self-consistent kinetic data on the interaction between site A pentadecapeptides and mAb SD6 Ž . Gomes et al, 2000 . In this paper, we present a definitive validation of our direct single-step SPR kinetic analysis and antigenicity ranking of small peptides. Using a panel of 44 FMDV site A-derived peptides and two anti-site A mAbs, we have obtained reliable kinetic parameters, good reproducibility and significant correlation Ž with data previously obtained by ELISA Verdaguer .…”

SPR: Uma nova ferramenta para biossensores

“…Os resultados demonstraram alta eficiência analítica e um potencial clínico no diagnóstico da hepatite A. Este dispositivo mostrou ser uma ferramenta poderosa para rastreamento clínico, de fácil uso, rápido e confiável. Outras aplicações dessa natureza podem ser encontradas na literatura [53][54][55][56][57][58] . A SPR também pode ser usada para monitorar interações proteína-proteína, ou proteína-ligante em tempo real e ainda permite a quantificação das espécies que estão interagindo.…”